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Figure 3 | Journal of Nanobiotechnology

Figure 3

From: Interaction of nanoparticles with proteins: relation to bio-reactivity of the nanoparticle

Figure 3

Schematic representation of the commonly used strategy to isolate and identify surface adsorbed proteins, when nanoparticles interact with complex protein mixtures. (A) Incubation of NP with protein solutions results in adsorption of protein onto the NP surface. Protein concentration may affect the amount and identity of proteins adsorbed on the presented NP surface. (B) Centrifugation for removal of unbound proteins followed by repeated washing of the NP-protein pellet is important for isolation of the “hard protein corona”. (C) Isolation of the NP-PC can be achieved by elution of the adsorbed proteins using denaturing agents such as Laemmli buffer which contains sodium dodecyl sulphate and 2-mercaptoethanol that facilitate the overall desorption of the proteins. (D) The desorbed proteins can thus be separated using one or two dimensional gel electrophoresis. (E) Separated protein bands of interest can further be subjected to tryptic digestion and can be subsequently identified using mass spectrometric methods.

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